Fig. 3: Detection of oxalyl-AMP as the intermediate in the synthesis of ODAP by LsAAE3.
From: Reply to: LsBOS utilizes oxalyl-CoA produced by LsAAE3 to synthesize β-ODAP in grass pea
a Extracted ion chromatograms of incubations of 100 mM Tris pH 8.0, 2 mM DTT, 5 mM ATP, 10 mM MgCl2, 0.3 mM sodium oxalate, 0.1 µM CoA with no enzyme, with LsAAE3, with LsAAE3 and desCoA replacing CoA, with LsAAE3, CoA and DAP. Black arrows indicate peak (m/z 418.0406) for oxalyl-AMP present in reactions with LsAAE3 and CoA or desCoA, which is absent when DAP is included as the oxalyl acceptor in the reaction. b Accurate mass determination with selected ion monitoring for peak m/z 418.0406. The calculated m/z was 418.04055, the observed was 418.0404 (indicated by black arrow) with an error of −0.36 ppm. c MS2 of selected ion m/z 418.0406. The calculated m/z for AMP in negative ion mode is 346.05581, the observed was 346.0551 with an error of −2.05 ppm. Source data are provided as a Source Data file.
